Proteomic Analysis Reveals Aberrant O-GlcNAcylation of Extracellular Proteins from Breast Cancer Cell Secretion

Cancer Genomics - Proteomics

JISNUSON SVASTI³  CHANTRAGAN SRISOMSAP³  PUKKAVADEE NETSIRISAWAN¹  VORARATT CHAMPATTANACHAI³  DARANEE CHOKCHAICHAMNANKIT² 

[1] pplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandpplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandpplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailand;aboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailand;pplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailandpplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandpplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailandpplied Biological Sciences Program, Chulabhorn Graduate Institute, Laksi, Bangkok, Thailandaboratory of Biochemistry, Chulabhorn Research Institute, Laksi, Bangkok, Thailand

关键词: Breast cancer;    extracellular proteins;    heat-shock 70 kDa;    O-GlcNAcylation;    transitional endoplasmic reticulum ATPase;   

DOI  :  

来源: Delinasios GJ CO

【 摘 要 】

Background: O-GlcNAcylation is a unique intracellular protein modification; however, few extracellular O-GlcNAc-modified proteins have been discovered. We have previously demonstrated that many cellular proteins were aberrant in O-GlcNAcylation in breast cancer tissues. In the present study, therefore, we investigated whether O-GlcNAc-modified proteins were abnormally secreted from breast cancer cells. Materials and Methods: Intracellular and extracellular proteins were prepared from cell lysates of breast cancer cells (MCF-7 and MDA-MB-231) and normal breast cells (HMEC) and from their serum-free media (SFM), respectively. O-GlcNAcylation level was examined by immunoblotting. O-GlcNAc-Modified proteins were identified using two-dimensional gel electrophoresis and Liquid Chromatography-tandem Mass Spectrometry. Results: O-GlcNAcylation level was significantly increased in the extracellular compartment of both types of cancer cells compared to normal cells. Interestingly, O-GlcNAc patterns differed between intracellular and extracellular proteins. Proteomic analysis revealed that many O-GlcNAc spots in MCF-7 secretions were abnormally increased in comparison to those in HMEC secretions. Among these, transitional endoplasmic reticulum ATPase (TER ATPase) and heat-shock 70 kDa (HSP70) were confirmed to be O-GlcNAc-modified. The levels of O-GlcNAc-HSP70 and O-GlcNAc-TER ATPase were higher in SFM from MCF-7 cells than in that from HMEC. Conclusion: O-GlcNAcomic study of the extracellular compartments reveals aberrant O-GlcNAc-secreted proteins, which may be of interest as potential biomarkers in breast cancer.

【 授权许可】

Unknown